ArticlePloS one2017
The widely used Nicotiana benthamiana 16c line has an unusual T-DNA integration pattern including a transposon sequence.
Article in PloS one, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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13 citing papers in PubMed, 30 citations in OpenAlex.
- Exogenous 24nt siRNAs induce AGO4A-dependent silencing via promoter DNA methylation and H3K9me2 deposition.Frontiers in plant science · 2026Article
- A multi-omic Nicotiana benthamiana resource for fundamental research and biotechnology.Nature plants · 2023Article
- Issue when expressing a recombinant protein under the control of pFrontiers in plant science · 2023Article
- A beneficial fungal root endophyte triggers systemic RNA silencing and DNA methylation of a host reporter gene.RNA biology · 2023Article
- Small RNA-seq dataset of wild type and 16CData in brief · 2022Article
- Magnetofection approach for the transformation of okra using green iron nanoparticles.Scientific reports · 2022Article
- Horizontal gene transfer from genetically modified plants - Regulatory considerations.Frontiers in bioengineering and biotechnology · 2022Article
- Systemic GFP silencing is associated with high transgene expression in Nicotiana benthamiana.PloS one · 2021Article
- Are the current gRNA ranking prediction algorithms useful for genome editing in plants?PloS one · 2020Article
- High-Pressure-Sprayed Double Stranded RNA Does Not Induce RNA Interference of a Reporter Gene.Frontiers in plant science · 2020Article
- Combining a Simple Method for DNA/RNA/Protein Co-Purification and Arabidopsis Protoplast Assay to Facilitate Viroid Research.Viruses · 2019Article
- The Rapid Methylation of T-DNAs UponFrontiers in plant science · 2019Article
- An Insight into T-DNA Integration Events in Medicago sativa.International journal of molecular sciences · 2017Article
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Authors and funding
6 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Nicotiana benthamiana is employed around the world for many types of research and one transgenic line has been used more extensively than any other. This line, 16c, expresses the Aequorea victoria green fluorescent protein (GFP), highly and constitutively, and has been a major resource for visualising the mobility and actions of small RNAs. Insights into the mechanisms studied at a molecular level in N. benthamiana 16c are likely to be deeper and more accurate with a greater knowledge of the GFP gene integration site. Therefore, using next generation sequencing, genome mapping and local alignment, we identified the location and characteristics of the integrated T-DNA. As suggested from previous molecular hybridisation and inheritance data, the transgenic line contains a single GFP-expressing locus. However, the GFP coding sequence differs from that originally reported. Furthermore, a 3.2 kb portion of a transposon, appears to have co-integrated with the T-DNA. The location of the integration mapped to a region of the genome represented by Nbv0.5scaffold4905 in the www.benthgenome.com assembly, and with less integrity to Niben101Scf03641 in the www.solgenomics.net assembly. The transposon is not endogenous to laboratory strains of N. benthamiana or Agrobacterium tumefaciens strain GV3101 (MP90), which was reportedly used in the generation of line 16c. However, it is present in the popular LBA4404 strain. The integrated transposon sequence includes its 5' terminal repeat and a transposase gene, and is immediately adjacent to the GFP gene. This unexpected genetic arrangement may contribute to the characteristics that have made the 16c line such a popular research tool and alerts researchers, taking transgenic plants to commercial release, to be aware of this genomic hitchhiker.
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